阿江榄仁酸由AMPK/mTOR/HO-1信号通路调控自噬对糖尿病视网膜病变影响  

Arjunolic acid protects diabetic retinopathy by regulating autophagy through the AMPK/mTOR/HO-1 signaling pathway

作　　者：蒋晨[1] 万新娟[1] 王绍飞[1] 王晓虹[1] 丁琳[1] JIANG Chen;WAN Xinjuan;WANG Shaofei(Department of Ophthalmology,People’s Hospital of Xinjiang UyguRAutonomous Region,Xinjiang,Urumqi 830002,China)

机构地区：[1]新疆维吾尔自治区人民医院眼科,乌鲁木齐市830002

出　　处：《河北医药》2024年第2期171-175,共5页Hebei Medical Journal

基　　金：新疆维吾尔自治区自然科学基金资助项目(编号:2021D01C150)。

摘　　要：目的探讨阿江榄仁酸(arjunolic acid,AA)对糖尿病视网膜病变(diabetic retinopathy,DR)大鼠视网膜细胞自噬及AMPK/mTOR/HO-1信号通路的影响。方法以健康SD大鼠为研究对象,构建链脲佐菌素(STZ)诱导的糖尿病大鼠模型,随机分为对照组(Con)组、模型(STZ)组、AA低剂量(AAL,10 mg/kg)组和AA高剂量(AAH,10 mg/kg)组。连续给药10周后,HE染色检测视网膜组织病理结构;qRT-PCR检测视网膜组织白介素(IL)-1β、IL-6和线粒体丙酮酸转运载体(MPC)-1的mRNA表达;二氢乙锭(DHE)染色评估视网膜组织ROS产生;Western blot检测自噬和AMPK/mTOR/HO-1信号通路相关蛋白表达。结果与Con组比较,STZ组大鼠视网膜出现肿胀和空泡样变化等病理变化,中央视网膜ONL层厚度和细胞核计数明显降低(P<0.01);IL-1β、IL-6和MCP-1的mRNA水平显著增高(P<0.05);视网膜外核层(ONL)、内核层(INL)和神经节细胞层(GCL)中ROS产生增加(P<0.01);LC3II/I比率、HO-1和p-AMPK/AMPK蛋白表达显著降低,p62和p-mTOR/mTOR表达升高(P<0.01)。与STZ组比较,AAL和AAH组大鼠视网膜ONL厚度和细胞核计数逐渐升高,结构相对规整(P<0.05);AAH组IL-1β、IL-6和MCP-1的mRNA表达明显降低(P<0.05);视网膜ONL、INL和GCL中ROS产生逐渐降低(P<0.01);LC3II/I比率、p-AMPK/AMPK和HO-1表达逐渐升高,p62和p-mTOR/mTOR表达逐渐降低(P<0.01)。结论阿江榄仁酸可能是治疗DR的候选药物,可能机制为通过AMPK/mTOR/HO-1调节的自噬途径保护视网膜细胞免受STZ诱导的氧化应激和炎症损伤。Objective To investigate the effects of arjunolic acid(AA)on autophagy and the AMP-activated protein kinase(AMPK)/mechanistic target of rapamycin(mTOR)/heme oxygenase-1(HO-1)signaling pathway in retinal cells of rats with diabetic retinopathy(DR).Methods A streptozotocin(STZ)-induced diabetic rat model was constructed in healthy Sprague-Dawley(SD)rats.Rats were randomly divided into control(Con)group,model(STZ)group,low-dose AA(AAL,10mg/kg)group and high-dose AA(AAH,10mg/kg)group.After 10 weeks of the continuous intervention,H&E staining was performed to detect retinal histopathological structures.The quantitative reverse transcriptase PCR(qRT-PCR)was performed to detect mRNA expressions of interleukin(IL)-1β,IL-6 and mitochondrial pyruvate transporter (MPC)-1 in retinal tissues.Dihydroethidium(DHE)staining was performed to assess the production of reactive oxygen species(ROS)in retinal tissues.Western blot was performed to detect autophagy proteins and those in the AMPK/mTOR/HO-1 signaling pathway.Results Compared with those of the Con group,rats in STZ group presented pathological changes like Retinal swelling and vacuolar degeneration,significantly lower thickness of the outer nuclear layer (ONL)of the retina,nuclear counting,light chain 3/1(LC3Ⅱ/Ⅰ)ratio and protein levels of HO-1 and p-AMPK/AMPK,but significantly higher mRNA levels of IL-1β,IL-6 and MCP-1,ROS levels in the ONL,inneRnuclear layer (INL)and ganglion cell layer (GCL),and protein levels of p62 and p-mTOR/mTOR(P<0.01).Compared with those of STZ group,ONL thickness and nucleus count of rats in the AAL group and AAH group were significantly higher (P<0.05),with a relatively intact retinal structure.Compared with those of AAL group,rats in the AAH group presented significantly lower mRNA levels of IL-1β,IL-6 and MCP-1,ROS levels in the ONL,INL and GCL,and protein levels of p62 and p-mTOR/mTOR,but significantly higher LC3Ⅱ/Ⅰratio and protein levels of HO-1 and p-AMPK/AMPK(P<0.05 oRP<0.01).Conclusion AA may be a promising candidate for the tr

关 键 词：阿江榄仁酸 糖尿病视网膜病变 AMPK/mTOR/HO-1通路 自噬 

分 类 号：R744.1[医药卫生—神经病学与精神病学]